Thesis Title

Identification and Analysis of a Putative Cell Surface Receptor For C. Elegans Collagen Type Iv Using Bioinformatics Tools and Rna Interference Mechanism

Date of Graduation

Fall 2006

Degree

Master of Science in Cell and Molecular Biology

Department

Biomedical Sciences

Committee Chair

Colette Witkowski

Keywords

Collage IV, RNAi, bioinformatics tools, microinjection, Meckel syndrome

Subject Categories

Medical Molecular Biology

Abstract

Collagen IV is an abundant basement membrane protein which forms a polygonal net type structure with the lateral association of its C-terminal and N-terminal domains providing support and stability to the basement membrane against mechanical forces. Collagen IV is involved in human genetic diseases such as Good Pasture syndrome and Alport’s syndrome demonstrating the importance of functional collagen IV protein in the basement membrane. The current research attempted to identify a collagen IV cell surface receptor in C. elegans utilizing different Bioinformatics tools and methods like BLAST, PSI-BLAST, ELM, clustral multiple alignment, Motif scan, and wormbase. These protein databases and search strategies have identified candidate genes F35D2.3, C37C3.7, T25F10.3 encoded proteins with potential domains required to recognize collagen IV and to function as a cell surface receptor. RNA interference was used as a gene silence technique to test whether the putative proteins might function as cell surface receptors. Microinjection was used as a standard method to deliver the dsRNA for RNAi mechanism to produce phenocopies. These putative proteins are being screened with RNAi for phenotypes similar to collagen IV mutants. No significant percent of embryotic lethality was observed from RNAi mediated identified candidate genes suggesting that the putative proteins identified were not the collagen IV cell surface receptors. Hypothetical protein F35D2.3 showed different abnormalities in the RNAi screens and was found to have amino acid sequence similarity and abnormalities to Meckel protein in humans.

Copyright

© Srilatha Nalluri

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Dissertation/Thesis

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